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Construction and evaluation of a novel protein mechanosensor
Kolomazníková, Veronika ; Rösel, Daniel (advisor) ; Novotný, Ivan (referee)
The protein p130Cas (human ortholog BCAR1) is a major substrate for phosphorylation by the Src family kinase and plays a central role in oncogenic transformation. Increased level of BCAR1 correlates with primary tumour growth and cancer progression. Localized to focal adhesion, p130Cas serves as a mechanosensor and mediates key interactions with the extracellular environment. The structure of p130Cas is crucial for its function, mainly the anchoring domains SH3 and CCH, together with the substrate domain which is extended when under tension. This Master's thesis presents a newly developer FRET mechanosensor based on the structure of p130Cas. The sensor utilizes the anchoring domains of p130Cas for proper localization to focal adhesions, where it can detect tension in living cells. Key words: p130CAS, FRET, focal adhesions, mechanosensing
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Construction and evaluation of a novel protein mechanosensor
Kolomazníková, Veronika ; Rösel, Daniel (advisor) ; Novotný, Ivan (referee)
The protein p130Cas (human ortholog BCAR1) is a major substrate for phosphorylation by the Src family kinase and plays a central role in oncogenic transformation. Increased level of BCAR1 correlates with primary tumour growth and cancer progression. Localized to focal adhesion, p130Cas serves as a mechanosensor and mediates key interactions with the extracellular environment. The structure of p130Cas is crucial for its function, mainly the anchoring domains SH3 and CCH, together with the substrate domain which is extended when under tension. This Master's thesis presents a newly developer FRET mechanosensor based on the structure of p130Cas. The sensor utilizes the anchoring domains of p130Cas for proper localization to focal adhesions, where it can detect tension in living cells. Key words: p130CAS, FRET, focal adhesions, mechanosensing
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Genetically encoded biosensors of cellular tension and their use in cellular biology
Pelantová, Markéta ; Rösel, Daniel (advisor) ; Lánský, Zdeněk (referee)
1 Abstract and key words Mechanical forces have great impact on the life of cells. They influence cell proliferation, migration or differentiation and defects in cellular mechanosensing were reported to be the cause of various diseases, such as deafness, atherosclerosis or cancer. However, mechanisms of mechanical sensing are not thoroughly examined and not many tools for doing such research are available. Genetically encoded FRET-based biosensors are one of the existing methods for studying transfer of mechanical signal in cells. It is a non-invasive method allowing to observe changes in mechanical tension across proteins in living cells. In this thesis, different types of existing genetically encoded FRET-based tension biosensors are introduced together with the process of their development and knowledge gained by their use in research. Key words: mechanical force, mechanosensing, FRET, tension sensor, biosensor development
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